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Related Experiment Videos

Polyubiquitin-regulated DsRed marker for transgenic insects.

A M Handler1, R A Harrell

  • 1US Department of Agriculture, Gainesville, FL 32608, USA. handler@nersp.nerdc.ufl.edu

Biotechniques
|October 30, 2001
PubMed
Summary

Researchers developed a new fluorescent marker, DsRed, for insect genetic transformation. DsRed allows for clear detection of multiple transgenes and gene expression when used with Green Fluorescent Protein (GFP) in Drosophila melanogaster.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Genetic transformation in insects often relies on dominant markers like Green Fluorescent Protein (GFP).
  • Detecting multiple transgenes or monitoring gene expression alongside GFP requires distinct, co-expressed visible markers.
  • Existing markers may lack spectral separation, hindering unambiguous detection when combined with GFP.

Purpose of the Study:

  • To evaluate the DsRed fluorescent protein as a selectable marker for piggyBac transformation in Drosophila melanogaster.
  • To assess DsRed's utility as a reporter for gene expression studies, particularly when co-expressed with GFP.
  • To establish a reliable method for distinguishing multiple fluorescent markers in genetically modified insects.

Main Methods:

Related Experiment Videos

  • Utilized the DsRed fluorescent protein as a marker in piggyBac-mediated transformation of Drosophila melanogaster.
  • Co-expressed DsRed with Green Fluorescent Protein (GFP) and its variants in transgenic flies.
  • Employed specific filter sets (Texas Red) to differentiate DsRed fluorescence from GFP and autofluorescence.

    • Main Results:

    • DsRed-marked transformants were identified with high frequency throughout development.
    • DsRed exhibited brighter fluorescence compared to Enhanced Green Fluorescent Protein (EGFP).
    • Using a Texas Red filter set allowed unambiguous detection of DsRed, even when co-expressed with EGFP, by eliminating EGFP and autofluorescence.

    Conclusions:

    • DsRed is an efficient fluorescent marker for selecting transformant insects.
    • DsRed serves as a valuable reporter for gene expression studies in insects, especially when combined with GFP.
    • The spectral distinctiveness of DsRed enables multiplexing fluorescent markers in genetic studies.